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Objective@#To investigate the effects and the potential mechanism of shikonin on rat random flaps.@*Methods@#Seventy-two wistar male rats in grade SPF were randomly divided into negative control group, tetramethylpyrazine group (TMP group) or shikonin treatment group. The random skin flap sized 8 cm×2 cm, with a cephalic based pedicle, was performed on the back of the rat. Each group was administered accordingly by intraperitoneal injection once per day for 7 days: shikonin treatment group (1 mg/kg), TMP group (10 mg/kg) and control group (1 ml/kg). Morphological changes of skin flaps were observed by HE staining. The positive expression of iNOS and COX-2 in skin flap tissues after operation were detected by immunofluorescence staining. The serum contents of TNF-α and IL-6 were detected by ELISA.@*Results@#Inflammatory cell infiltration and inflammatory reaction of flap was more severe in control group at different time points after operation. The number of inflammatory cells in shikonin treatment group and TMP group were significantly decreased, compared with controls (P<0.01). The decrease of the inflammatory cell numbers in shikonin treatment group was more significant. The proliferation of granulation tissue and fibroblast in skin flap was obvious, and the survival rates of the skin flap were significantly increased in shikonin treatment group and TMP group (both P<0.01). The numbers of iNOS or COX-2 positive cells in shikonin treatment group and TMP group were significantly decreased, when compared with controls (both P<0.01). Compared with TMP group, the numbers of iNOS and COX-2 positive cells in shikonin treatment group were significantly decreased (P<0.01 and P<0.05, respectively)in early period after operation. Compared with the control group, serum levels of TNF-α and IL-6 in shikonin treatment group and TMP group were significantly decreased (all P<0.01). The shikonin treatment group developed more significant reduce.@*Conclusions@#Shikonin can significantly improve inflammatory response of skin flap in rats, which might be related to inhibiting the expression of iNOS and COX-2, decreasing serum levels of TNF-α and IL-6 , and reducing the release of inflammatory cytokines .
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Biochemistry is an essential course for medical students,which updates fast,develops quickly and has strong characteristic of experimentation.Opening experiment is a new open type experimental teaching mode and students are regarded as main body.Opening experiment is helpful to stimulating activeness and cultivating innovative consciousness of students as an effective supplement for traditional experimental teaching.This kind of teaching mode is in conformance with experimental teaching reform in university and is an effective way of cultivating medical science talented men.
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Aim A UVA-induced apoptotic model of HaCaT cells was established to investigate the impact of UVA on c-jun/cyclooxygenase-2(COX-2)and explore related molecular mechanism of the polypeptide from Chlamys farreri(PCF)protecting HaCaT cells from UVA-induced apoptosis.Methods Cells were divided into five groups:control group,UVA model group,UVA+5.69 mmol?L-1 PCF group,UVA+2.84 mmol?L-1 PCF group,UVA+1.42 mmol?L-1 PCF group.Expression level of c-jun was assayed by Real-Time PCR and Western blot.RT-PCR and Western blot analysis were used to determine the mRNA and protein levels of COX-2.Using agarose gel electrophoresis,the effects of PCF and COX-2 inhibitor celecoxib on UVA-induced apoptosis were also investigated.Results PCF and celecoxib had inhibitory effect on 8 J?cm-2 UVA-induced apoptosis of HaCaT cells.COX-2 mRNA and protein levels increased after UVA radiation and the discrepancy was significant compared with control group(P